The researchers reported last October that full-length Shh protein present on the cell membrane identified cells with cancer stem cell (CSC) features implicated in NSCLC. To target these perpetrator CSCs and disable them, they developed therapeutic antibodies against the carboxy (C)-terminus of Shh and characterized their pre-clinical activities in cancer cells and mice. This is the first reportof a therapeutic and inhibitory antibody designed to specifically target the carboxy-terminus of Shh. Lead author Tolani played a pivotal role in guiding the team, and collaborated closely with Jablons throughout the project. The paper was featured on the cover of the March 6th issue of the journal Oncotarget.

Schematic of experimental procedures for generation of novel anti-Sonic Hedgehog (Shh) candidate therapeutic antibodies directed at the carboxy (C)-terminal. (A) Two synthetic peptide mimics of Shh (AA 247-264 and AA 448-462) were conjugated to carrier protein KLH and used as the immunogen. (B) Dual administration of both peptides was used to elicit an immune response in Sp2/0-Ag1 mice. (C) B cells from immunized murine lymph nodes were isolated and fused to myeloma cells for hybridoma generation. (D) Protein-based screening analyses guided the selection of the best hybridoma clones followed by one round of sub-cloning to isolate monoclonal antibody-producing hybridoma cells (E) Large-scale preparations of purified antibodies from hybdridomas were used for in vitro, in vivo, and ex vivo investigations.